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. 2019 Jun 13;105(2):508–518. doi: 10.3324/haematol.2018.212118

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Figure 4. — Readthrough agents (RTA) reduced intracellular accumulation of FVIII in transfected Chinese hamster ovary (CHO) cells. Representative images of the immunodetection of FVIII heavy chain in cell homogenates of transiently transfected CHO cells. (A) Western blot of CHO cells transfected with either wild-type (WT) BDD-FVIII (166.19 kDa), the p.Q462X (52.8 kDa), p.Q1764X (98.51 kDa) (top panel), p.W2015X (127.9 kDa), or the p.R1822X (105.24 kDa) variants (bottom panel). The intracellular accumulation of BDD-FVIII or the truncated proteins before and after RTA-treatment is shown. (B) Densitometry of the bands in (A). CT: Untreated cells; GN50: 50 mg geneticin/mL; GN100: 100 mg geneticin/mL; GT50: 50 mg gentamicin/mL; GT100: 100 mg gentamicin/mL; PTC: PTC124 10 mM; RTC13 and RTC14: 10 mM; Mock: untransfected cells; MW: Pageruler Plus prestained protein ladder; Refacto®: recombinant human BDD-FVIII (Pfizer); LR: loading reference used for densitometry normalization.